基于近红外免疫层析技术食源性诺如病毒快速检测方法研究  

Rapid Detection Method of Foodborne Norovirus Based on Near-infrared Immunochromatography

作　　者：张捷[1,2] 王琳 霍江莲[4] 杨向莹[1,2] 畅晓晖[1,2] 李小林[1,2] 张园[5] 陈广全[1,2] 

机构地区：[1]北京出入境检验检疫局检验检疫技术中心,北京100026 [2]出入境食品安全检测北京市重点实验室,北京100026 [3]国际检验检疫标准与技术法规研究中心,北京100028 [4]中国合格评定国家认可中心,北京100162 [5]天津出入境检验检疫局工业产品安全技术中心,天津300308

出　　处：《黑龙江医学》2017年第7期691-694,共4页Heilongjiang Medical Journal

基　　金：质检公益性行业科研专项项目(201410049);国家质检总局科技计划项目(2015IK011);(2013IK144)

摘　　要：目的建立一种快速、灵敏检测食源性GⅡ型诺如病毒的近红外免疫层析法。方法采用低噪声激发式荧光染料标记鼠抗诺如病毒单克隆抗体,将诺如病毒多克隆抗体和羊抗鸡Ig Y多克隆抗体喷涂于硝酸纤维膜上分别作为检测线和质控线,制备靶向于诺如病毒衣壳蛋白的免疫层析试纸条,研制检测诺如病毒的近红外免疫层析试纸条。结果采用便携式低噪声激发式荧光扫描仪分别扫描质控线和检测线,以检测线荧光强度检测值实现样本的定性检测,在45 min内即可完成检测。该试纸条与肠道病毒EV71、腺病毒、轮状病毒、甲肝病毒均无交叉反应。结论 GⅡ型诺如病毒的近红外免疫层析法具有良好的特异性和较高的灵敏度。通过效果评价试验发现,与荧光RT-PCR、胶体金法相比较,所建立的近红外荧光法检测时间最短,检测限低于胶体金法。因此,近红外免疫层析法可用于食品中GⅡ型诺如病毒的高效检测,为食品安全监管提供可靠的技术支持。Objective To establish a rapid,sensitive detection of Group GⅡ norovirus near-infrared immunochromatography.Methods Through using near-infrared fluorescence labeled murine monoclonal antibody against norovirus,norovirus polyclonal antibodies and goat anti-chicken Ig Y polyclonal antibody were sprayed on the nitrocellulose membrane. Then those were used as test line and control line and developed to detect norovirus NIR immunochromatographic strip and supporting reference materials. Results Near-infrared fluorescence was completed in 45 minutes and no cross reaction with EV71 virus,adenovirus,rotavirus and hepatitis A virus. Conclusion NIR immunoassay for the detection of Group GⅡnorovirus has good specificity and high sensitivity. Evaluation by the test found,near-infrared fluorescence detection with RT-PCR and colloidal gold wears comparison,the established minimum detection time. Near-infrared fluorescence detection limit is lower than the colloidal gold. Thus,the near-infrared immunoassay can be used to efficiently detect norovirus in food,to provide reliable technical support for food safety supervision.

关 键 词：诺如病毒 快速检测 近红外免疫层析 灵敏性 特异性 

分 类 号：R446.1[医药卫生—诊断学]